【摘要】：樹(shù)突狀細(xì)胞(dendritic cell, DC)是已知體內(nèi)功能最強(qiáng),唯一能活化初始型T 細(xì)胞的專職抗原遞呈細(xì)胞(antigen presenting cell, APC),其數(shù)量、成熟狀態(tài)可決定機(jī)體免疫狀態(tài),故對(duì)其深入的研究具有重要理論和實(shí)際意義。DC 具有體外可操作性,因此獲取一定量有功能的DC 是深入研究其生物特性的關(guān)鍵。 本研究目的為在體外建立一個(gè)穩(wěn)定的從人外周血單核細(xì)胞誘導(dǎo)未成熟及成熟DC 的培養(yǎng)方法,經(jīng)流式細(xì)胞術(shù)檢測(cè)細(xì)胞表面分子的表達(dá)情況并應(yīng)用MTT 法對(duì)培養(yǎng)細(xì)胞的生物學(xué)功能進(jìn)行測(cè)定,從而確保獲得具有一定形態(tài)和功能特征的DC,為進(jìn)一步探討DC 在免疫耐受及抗腫瘤方面作用奠定基礎(chǔ)。 研究結(jié)果:本研究建立了誘導(dǎo)外周血單核細(xì)胞獲得不同發(fā)育階段DC的方法,獲得較大量,純度較高的CD83+DC。與以往報(bào)道有所不同,經(jīng)流式細(xì)胞術(shù)檢測(cè)高表達(dá)CD83分子的DC仍有CD14分子的表達(dá)。經(jīng)過(guò)再次混合淋巴細(xì)胞反應(yīng)發(fā)現(xiàn)具有中度表達(dá)共刺激分子的未成熟DC在體外可以誘導(dǎo)同一來(lái)源的T細(xì)胞免疫低應(yīng)答。 通過(guò)本研究我們建立了穩(wěn)定的體外DC 培養(yǎng)體系,為移植免疫耐受研究和抗腫瘤研究深入發(fā)展提供了理想的基因轉(zhuǎn)移靶細(xì)胞。
[Abstract]:Dendritic cell (dendritic cell, DC) is known to have the strongest function in vivo. The only specialized antigen-presenting cell (antigen presenting cell, APC),) that activates the initial T cell is the number of (antigen presenting cell, APC), and the mature state determines the immune state of the body. Therefore, the in-depth study of DC has important theoretical and practical significance. DC can be operated in vitro, so obtaining a certain amount of functional DC is the key to further study its biological characteristics. The aim of this study was to establish a stable culture method for inducing immature and mature DC from human peripheral blood monocytes in vitro. The expression of cell surface molecules was detected by flow cytometry and the biological function of cultured cells was determined by MTT assay. So as to ensure that DC, with certain morphological and functional characteristics lay a foundation for further study of the role of DC in immune tolerance and antitumor. Results: in this study, we established a method to induce peripheral blood monocytes to obtain DC at different developmental stages. A large number of CD83 DC. with high purity were obtained. Different from previous reports, DC with high expression of CD83 still expressed CD14 by flow cytometry. After remixed lymphocyte reaction it was found that immature DC with moderate expression of costimulatory molecules could induce low immune response of T cells from the same source in vitro. Through this study, we established a stable culture system of DC in vitro, which provides an ideal target cell for gene transfer for the further development of transplantation immune tolerance and antitumor research.
【學(xué)位授予單位】：吉林大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2005
【分類號(hào)】：R392

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